Work

Abstract 1997 Proteomic and structural analysis of the oncogenic EWS::FLI1 spliceosomal interactome

Year: 2024

Type: conference-abstract

Abstract: The EWS::FLI1 gene fusion is implicated as a source of oncogenic activity in the majority of Ewing sarcoma (EwS) cases (>70%).EWS::FLI1 is composed of RNA-binding protein EWS (EWS) and ETS transcription factor Friend leukemia integration 1 (FLI1).It is thought that EWS::FLI1 and EWS participate in mRNA processing and spliceosomal activities, however, the structural interaction mechanisms are unknown.Proteomic experiments show spliceosomal proteins are present in both EWS and EWS::FLI1 interactomes, including proline-rich-region-binding spliceosomal peptidyl-prolyl isomerases (PPIs).EWS::FLI1 contains a low complexity domain (LCD), enriched in proline residues, imparts intrinsic structural disorder and acts as a multivalent binding region.Here we used modern proximity labelling techniques that exploit a biotin ligase fused to EWS and EWS::FLI1 to identifying their interactomes.This approach is particularly well suited for intrinsically disordered proteins since it does not rely on binding affinities but rather the physical distance between interacting proteins.From these experiments we identified specific and unique interactions between EWS and EWS::FLI1 and PPI proteins that we hypothesized were mediated by the LCD (EWSLCD) common to both proteins.We employed NMR to provide insight into the interaction between PPIL1 and the EWSLCD and identified the binding sites on both proteins.Specifically, our studies will yield actionable insights regarding the protein-protein interfaces in EWS::FLI1 that can be targeted to attenuate its oncogenic activity.Defining the interaction interfaces of EWS::FLI1 is central to understanding its pathophysiology and importantly, identification of potential therapeutic targets.Combining novel discovery proteomics with cutting-edge NMR approaches will provide unparalleled insight into the transformative mechanics of EWS::FLI1.DSL is the Shohet Family Fund for Ewing Sarcoma

Source: Journal of Biological Chemistry

DOI: 10.1016/j.jbc.2024.107022

Authors: George Parra, David Libich, Antoine Baudin, Susan Weintraub, Bernard Fongang

Institution: The University of Texas at San Antonio Health Science Center

Language: English

Topic: Sarcoma Diagnosis and Treatment

Subfield: Pulmonary and Respiratory Medicine

Field: Medicine

Domain: Health Sciences

Open Access status: gold

Estimated APC paid: $2,800

Funder: National Institute of General Medical Sciences